Testes biopsy analysis and hormonal reproduction levels of pig tail macaque (Macaca nemestrina) injected with testosterone enanthate (TE) and depot medroxy progesterone acetate (DMPA)

(1)

Testes

biopsy analysis

and hormonal

reproduction

levels of pig

tail

macaque

(Macaca nemestrina) injected

with

testosterone

enanthate

(TE)

and depot

medroxy progesterone acetate

(DMPA)

Dadang

Kusmana*, O.

Soeradi+,

N. Moeloekt, N.

Suhana+, S.

Sri

Sukmaniah*, J. Supriatnax

Abstrak

Tuiuart penelitiatt

ini

adalalt untuk nrcnganalisa testis

lnsil

biopsi dan ntengetalrui kadar honnon reproduksi pada beruk (Macaca nentestritn)

jartan

tartg disurrtik kombinasi testosteron enantat (TE) dan depoî ntedroksiprogesteron asetat (DMPA). Sebaryak 6 bentk

iantan

berunrur antara

6

talum san4tai

8

talutr

digunakcn sebagai sampel. Masa adaptasi

3

bulan, kenudian dilakt*an l)ettyltttikatt TE dengart dosis 32 nry setiap nùtggu selama

6

nùtggu dan dilanjutkan setiap

3

nringgu sampai minggu ke 24, setlartgkart

pea'uûikan

DMPA dilakukan tiap

6

nùtrggtr dengan dosis 40 mg santpai ninggu

ke

18. Selain

itu

dilakukan pula Itonnott steroid. Biopsi testi.r clilakukan pada tninggu ke-30 dan ninggu ke-48. Penrbuatan sajian histologi testis dengan ntetode

ltarafut dart ltewarttaan hematoksilirt-eosirt (HE). Hasil penelitian nrcnwlukkan baltwa rata-rata volurne testis kanan

tlan

kiri

nt.crtgalami _{l)ulrffunan pada}nrasa perlakuan),aitu

dari

18.35 cnt3

+ 9.35

dan 19.02 an3

+

10.88 (ninggu ke 0) ntenjadi 6.70 cm3 + 3.80 dcut 7.02 cnt3 +

4.6I volunre terendah

pada nùnggu ke 2L Kemudian pada masa _{pemulihan akan meningkat kembali ),aitu}20.34 crrr3 + 7.87 dart 21.75 cnt3

+ 7.09

pada ninggu ke 48. Rata-rata dianteter tubulus setninferus dan skor spermatogenesis testis kanan

datt kiri pada akhir patgarulr TE dan DMPA (nùnggu ke 30) atlalah 0.13

nnt +

0.027 dan 0.13 nun + 0.026 serîa skor 5.08

+

2.67 dart 5.4

I +

2.5

l.

Pada

aklir

penelitian (rnasa pentulihan, nringgu ke 48) diameter tubulus nrenjadi 0.18 mm + 0.029 dan 0.18 mm + 0.026 scrta .rkor spennatogenesis 7.51

+

2.14 dan 7.57

+

1.59.

Di

sanqting im tubulus sentiniferus pada minggu ke 30 nrengalami lialirùsasi datt _{fibrosis. Kadar}lnnnon testosteron total, lestosteron bebas, dan estrogen unlunutta nteningkat tajam sampai minggu ke

6

tlan akan nretuu'utl kentbali tetapi

nnsih

lebih tinggi dibandingkan dengan kadar basalnl,a sarnpai mùrygu ke 30. Pada nnsa pemulilntt kadar estradiol nteningkat tajant kenr.bali santpai akhit'penelitian. Kadar hormon FSH dan LH menurun sanrpai minggu kc-6, kenurcliart honnon FSH akan nrcningkat kentbali sanqtai akhir penelitian, sedangkan _{ltada kadar hormon LH penurunatt}tersebut tctal) dipcrt(thankan santpai akhù' penelitian.

Dari

hasil penelitian disitttpulkan bahwa penyuntikan kontbinasi TE dan DMPA akan trtctrt1tertgarulti struktur lùstologi testis yang ditandai dengan pengecilan diameter tubulus seniniferus, hiaLinisasi, danfLbrosis serta merrylrunrbat pe rkembangatl spennatogetlesis. Gangguan yang

tejadi

pada testis tersebut lebih disebabkan oleh adanya hantbaîan

tcrlndap _{umlnn balik ltada poros hiltotalanrus-hiltofrsis-testis.}(Med J Jndones 2001; 10: 137-43)

Atrstract

Tltc objcctive o.f this suu[, i.ç

b

analyse tlrc testes biopsy and to know the lnrmone level reproduction of pig tail macaque (Macaca rt.crtrcstritut) injcctetl with tastosterone enantlnte (TE) and depot tnedro4progesterone acetate (DMPA). Six pig tail male macaques, age 6 - 8 ycars okl were usetl as sanrple. Three ntonths after adaptation period, each animal was injected

inta

muscularly with 32 mg TE cach we ek starting at week zero up to the sixth week. The tre(ltment was continued every 3 weeks after the sixth week up to the 24th vveck. 40 mg of DMPA was injected intranruscularly at week zero, and cotxtinued every week up to week i'8, Volume of the testes was takett every three weeks and bl.ood santples

for

examination of gonadotropin hormone and steroid hormone were taken at 6 week irûervals. Te.stes biopsy was perfonned ar week 30 and week 48. Preparation of testes histological slides were made using the

parffin

ntclhod anrl stained witlt hetnatorylin-Eosin (HE). The results of this study showed that both testes volume decreased

i.e.

18.35 cm'r + 9.35andl9.02cm.r

+

10.88(atweekzero)to6.70cm't

+3.80and7.02cmr

+4.61 (Iowestvolumeatweek2l). Inrecoveryperiod, thc testes vol.wne increased

to

20.34

cmr

+

7.87 and 21.75

cmt +

7.09. The diameter of the seminiferous tubules.and the icore oJ slrermatoge nesis (righ and left testes) at week 30 were 0.

l3

mm + 0.027 and 0.I 3 mm + 0.026 and score were 5.08 + 2.67 and 5.41 + 2.51. At week 48,_both diameter of seminferous.tubules and spermatogenesis score increased to become 0.18 mm

+

0.029 and 0.18 tnnx

+

0.026, and score were 7.51

+

2.14 and 7.57

+

1.59,

During this

period, hyalinization and _{fibrosis of}seminiftrous tubule occLtrred. By week

6,

the totaL testosterone, _freetestosterone, and estrogen hormone levels increased quite sharply and then decreased but still higher than base levels of hormone. In the recovery period, estrogen hormone increased significantly until the end of obsert'ation (week 48). FSH and LH hormone levels decreased

until

week

6, then

the

FSH hormone levels increased

until

the end

of obsertation,

while

the

LH hormone leveL

is

still lower than base level. Conclusion

of

this study

is

the injection

of

TE and DMPA combination wiIL

alter the

histological structure of

the

pig

tailed * Department of Biology, Facul.ty of Mathematics and

N a tural. S c i e nc e s, U niv ers ity of I ndo nes ia, J akar Ia, Indone s ia + _Dq)artnxent

of

Biology, Faculty of Medicine, IJniversity

of

Itdonesia, Jakarta, Indonesia

+

(2)

138

Kusmana et al Med J Indones

macaque testes i.e. decreasing the diameter of the seminiferous tubules, suppressing sPerrnatogenesis and hyaLinisation andfibrosis

of

seminiferous tubules. The datnages

of

this structure

are likely

caused

by

inhibition of feedback mechanism

of

hypothalamus-hypophysis-testes. (Med J Indones 2007

;

10: 137-43)

Keywords: Macaca nemeStrina, seminiferous tubule, spermatogenesis, gonadotropin hormone, steroid hormone.

The aim

of

male hormonal contraception

is to

suppress

spermatogenesis

process

via the

hypothalamus-hypophysis- testes axis.l-2 This method caused a decrease

in

gonadotropin

production,

and influenced

the

sper-matozoa

production and quality. The

admrnistration

of high

doses

of

testosteron enanthate

(TE) to

healthy

men through

intia

rnuscular

injection

induced

azoo-spermia and is

followed by lower

spermatozoa quality'3

Data

from

10

study

centers

in 7

cities concluded

that

administration

of

200

mg/week TE

Io

271healthy

and

fertile

men

caused

azoospermia

in

157

(65Vo)

of

the

volunteers after up

to

six

months

of

treatment.a

Combination

of

TE

and

DMPA

has already

been

ipvestigated

several

times

for

effectiveness.

Knuth

er

al

conducted a

trial on

12 volunteers

who

was

injected

with

200

mg

l9-nortestosterone

hexyloxyphehyl

propionate (19NT-HPP)

and 250

mg

DMPA,

azoospermia

were

achieved

in 6

men, while

4 men

achieved oligozoospermia

with

concentrations

<

2 x

106/mL, and

the remaining

2

men produced

only

a

single

sperm

per

ejaculate.s

In

contrast,

a

in

Indonesian men

given combination of

TE

and

DMPA, injection

demonstrated

higher rates

of

azoospermia.

In

a

trial

cqnducted

by

Pangkahila

on

10

volunteers,

azoospermia

were

achieved

in

all

volunteers

(10OVo),6

and one conducted

by

Moeloek

on

45 volunteers,

azoospermia was achieved in95.1Vo

of

the volunteers.T

The

studies above showed that

combination of

TE

and

DMPA

induced

azoospermia.

However.

the

exact

effect on

spermatogenesis

in

the testes

is still

unclear.

Therefore,

it

is

very

important

to

_.

understand

the

inhibition

of

spermatogenesis

by injection of TE

and

DMPA

combination.

Ohservations using testes

biopsy

from

men are considered unethical so therefore,

it is

important

to cârry

out the investigation using

animals

who evolutionafly

are close

with

humans.

The aim

of

this investigation

is to

perform an

analysis

of

testes

biopsy

and

horrnonal reproduction of pig

tail

macaque

testesinjected

with

combination

TE

and

DMPA.

METHODS

Animal

and

treatment

Six

male

pig tail

macaques (Macaca

nemestrina),

age

6-8

years

old

and

6-12

kg body weight

were

used

in

this

investigation.

They were

obtained

from

the Primate Research Center

of

Bogor Agriculnrre

Institute,

West

Java,

Indonesia.

Animals

were housed

in

cages

measuring

23x2lxl2

inches, maintained

at a

room

temperature

of

20o-2goc,

and

well

ventilated.

The standard

monkey-chow was

imported from

Seafoods

Enterprise

Co.

Ltd.

Thailand. The investigation

was

divided

into

three

periods

:

adaptation

period

for

3 months

from -12

week

to

zero

week;

treatment

period

for 6

months

from

zero week

to

24th

week; recovery

period

for

6 months

from

week 24

to

week 48.

Three

months

after the

adaptation

period,

each animal

was

injected

intra muscularly

with

32 mg

TE

each

week

starting

at

week zero

up

to

the sixth

week.

The

treatment was continued

every

3

weeks

after the

6th

week

up to the

24Ih

week.

Fourty mg

DMPA

was

injected intramuscularly

at week

zero, and continued

every week up to

week

18.

Testlcular volume

and

testicular

[iopsy

(3)

Hormone

measurements

Blood

samples

for

hormone

measurements

were

obtained

fiom

the

femoral vein.

Serum

was

frozen

at

-20oC

until assayed.

Serum steroids

(total

testosterone,

free

testosterone, and

estradiol) were

measured using

RIA kits

using

the

coat-A-Count

r25I

technique

as

described

by

the

producer (Diagnostic

Products

Corporation,

Los

Angeles,

CA

90045).

FSH

and

LH

were

measured

using double antibody

RIA kits

and

the

technique described

by the

producer (Diagnostic

Products

Corporation, Los

Angeles,

CA

90045).

RESULTS

Body weight

and

testes

volume

The

body weight

of

each

pig

tail

macaque at the week

0

befbre injection

averaged

9.10

kg

+

3.28.

During

treatment,

their body weight

increased.

By

the end

of

week 21,

the average

weight

rate reached

up

to 11.00

+

2.8'7. At the

recovery period

i.e. between week 30 to

week 48, the

body weight was

to

I 1.02

kg,

as shown

in Table

stable

around 9.82 kg

1.

The

average

volume

for

both right

and

left testes

at

the

week

0 were

18.35

cm'l

+

g.:S

and, 19.02

"rnt

+

10.88

respectively.

During

the

treatment

testes

volume

decreased and

the lowest

was at

the week

21;

the

average

volume

of right

and

left

testes

were

6.70

cmt

+

3.80

and

7.02

cm3 +

4.6l

respectively. During

the

recovery period,

each testes

volume

increased and

by

week

48, their

volumes

were larger

compared

to

the volume at week

0, i.e.

20.34

c-t

+

7.87 (right

testes)

and2l.l5

cm3

+

7.09

(left testes). See

Table

1.

Diameter

of

seminiferous

tubules

and

score of

spermatogenesis

After injection with

TE

and

DMPA at week 30, the

average

diameter

of

tubulus

seminiferous

were

0.

l3

mm +

0.027

(right

testes)

and 0.13

mm

10.026

(left

testes). During the

recovery

period, their

diameters

increased

(at week

48)

to

0.18 mm

+

0.029

(right

[image:3.612.46.484.420.693.2]

testes) and 0.18 mm

+

0.026

(left

testes). See Table 2.

Table l. Body weight and right and left testes volume before,?uring, and after injection with combination TE and DMPA

Weeks Body weight (kg)

+SD

Testes volume (cmr)

+SD

Right Left

-t2

0

3

6 9

t2

15

t8

2l

24

z7 30 JJ

36

39

42 45 48

8.28 + 3.07 9.10 + 3.28 9.48 + 3.35 10.4'1+3.64 10.25 + 3.24

10.38 + 3.21 10.67 + 3.17

10,81 + 3.04

I 1.00 + 2.87 10.95 +2.71 10;73 + 2.82

10.58 + 3.13 10.53 + 2.58

10.62 + 1.42

10.70 + 3.14 11.02 + 2.87 9.82 + 3.19 9.82 + 3.35

18.48 + 9.33

18,35 + 9.35 14.05 + 9.31

13.64 +7.00 9.42 + 3.89 8.22 + 3.65 9.63 + 5.17 11.65 + 4.26 6.70 + 3.80

8.07 + 4.08

8.39 + 6.22 8;72 + 3.95

12.98 +9.34

I 1.93 + 8.59 l7 .95 + 9.20

17 .05 + 9.20

16.47 + 6.38

20.40 +7.8'7

18.25

+

8.35 19.02 + 10.88

14.64

+

6.94

13.64

+ 9.25

10.01

+

6.39 8,61

+

4.50 12.94

+

8.89 9.07

+

5.62 7.02

+

4.61 8.92

+

5.78

8.07

+

4.82

10.31

+

4.52 13.40

+

0.87

13.21+

0.48 l'7.'75

+

2.41

18.80

+

9.93 17.68

+

6.52

(4)

140

Kusmana et al

Table

2.

Diameter of the seminiferous tubules and scores of the testes spermatogenesis before, during, and after injection with combination TE and DMPA

week-30

Med J Indones

Animal

no _{Diameter of seminiferous}tubules (mm)

+

SD

Scores of spermatogenesis

+SD

Right Right Left

I 2 3 4 5 6

0.10 + 0.026 0.14 + 0.019 0.14 + 0.023 0.08 + 0.015 0.16 + 0.015 0.14 + 0.023

0.10 + 0.025 0.14 + 0.024 0.14 + 0.017 0.08 + 0.012 0.15 + 0.017 0.14 + 0 020

1.66 + 1.21

6.08 + 1.63 '7.20 ₊_1.55

1.18 + 0.27 6.62 + 1.99

7.86 + 0.72

2.50 + 2.49 6.62 + 1.66 7.24 + 1.64 1.41 + 0.66 6.62 + 1.63

8.06 + 0.88

t

X+

SD

0.'76

0.13 + 0.027

0.75

0.1.3

+

2 67

30 50 5.08 + 2.67

32.45 5.41 + 2.51

week-48 Animal

no _{Diameter of seminiferous}tubules (mm)

+

SD

Scores of spermatogenesis

+SD

Right Right Left

1 2 3 4 5 6

0.12 + 0.031

0.18 + 0.029 0.19 + 0.025 0.20 + 0.037 0.19 + 0.027 0.21 + 0.023

0.13 + 0.037 0.18 + 0.027 0.18 + 0.017 0.16 + 0.025 0.20 + 0.023 0.21 + 0.028

3.20 + 2.31 8.86 + 1.04 8.90 + 1.17

6.36 + 3.75 9.06 + 1 .12

'7.86 ₊_0.'73

4.41 + 3.46 8.82 +

l.0l

8.64 + 1.64 6.02 + 3.98

8 70 + 1.39

8.50 + 0.96

t

X+

SD

1.09 1.06 45.06

0.18+0.029

0.18+0.026

7.51+2.14

45.42 7 .57 + 1.59

The average

score

of

spermatogenesis

in

the

seminiferous tubule

epithelium

at week

30

were

5.08

+

2.6'7

(right

testes)

and

5.41 + 2.51

(left

testes).

These

scores

indicated that

spermatogenic

cells

in

seminiferous

tubule do not contain

spermatozoa and

spermatid

cells

were

very few or

even

not

available.

Two of pig tail

macaques

were

in

deep

hyalinization

(see

Figure

1).

During

the recovery period,

histological

structure

of

seminiferous tubule epithelium

improved,

and these

are

shown through increasing

scores

at week

48, i.e.7

.51

+ 2.I4

(right

testes) and 7 .51

1

1.59

(left

testes). See

Table

2.

Steroid

hormones

Levels

of

total

testosterone hormone, free testosterone,

and estradiol at week

0 were

1.98

nglml +

1.0,2.69

pg/mL

+

1.54,

^nd

6.29 pglmL

+

7.48

respectively.

For

6

weeks

after

the first

injection,

the

levels ilrcreased

quite

sharply,

i.e.

8.52 nglrrJ-

+

1.52,

17.4

pg/ml

+

4.4I,

and

20J9

ng/ml

+

20.78 respectively.

After

week

6

the

average

level

of

the three

steroid

hormones

decreased.

Total

testosterone

levels

by

week 30

almost

reached

their

basal

levels

(week

0),

while free

testosterone

by

week 30 were quite

high

(4.54 pglmL

+

0.66)

compared

to

week

0,

and they

came

to their

basal levels

during

the recovery period

at

week

36.

Estradiol hormone levels

during

the

treatment

decreased

to

the lowest level

by

week

24

(2.43

pglmL

+

4.81)

and

from week 30

to

week

48

they

increased

drastically (10.18

pglml-

+

6.04

to

18.14

pg/mL

+

4;71).

See

Table

3.

Table

3.

Concentration ofthe steroid hormones serum before, during, and after TE and DMPA injection

Weeks Steroid hormones (n = 6)

Total

Free

Estradiol

testosterone testosterone

(pg/ml-) + SD (ne/ml-) +

SD

(pelml-) + SD

-12 0 6 12

l8

24 30 36 42 48

3.08 +

2.78

4.55 +3.26

1 .98 +

1.00

2.69 + 1.54 8.52 +

1.52

1'1.40 + 4.41 5.55 +

3.24

4.28 + 0.94 3.58

+2.27

5.09 + 1.59

4.31 +

2.15

10.75 + 4.42 2.96 +

2.18

4.54 + 0.66 3.12 +

4.47

1.27 + 0.32

3.24+336

3.43 +2.67

5.Q9 +

2.9'l

11.50 + 2.66

2.45

+

1.09 [image:4.612.53.496.120.406.2] [image:4.612.308.539.569.726.2]

(5)

[image:5.612.92.467.94.599.2]

Figure 1. Histol.ogical structure of the testes. A: Structure of the testes atweek-3}; B: Structure of the testes atweek-48.

(6)

142

Kusmana et al

Gonadotropin hormones

The

average

concentration

of gonadotropin

hormones

FSH

and

LH

at week

0

were

2.44 mIIJlmL

+

1.33 and

0.10

mIU/mL

+

1.33 and

0.10 mIUimL

t

0.00

respectively.

After injections

of TE

and

DMPA,

FSH

level

decreased

until

week 12 (28 mIU/mL

+

0.18).

After week

12

it

increased

gradually

and reached the highest

level

by

the

week

36

(4.91mIU +

6.48).

After

given

the

first injection

and

until

the study completed'

LH

hormone, however, decreased and

reached

the

lowest

level

at

the

week 6,

30,

and

48

i.e.

0.00

mIU/mL, 0.04 mIU/mL

+

0.07, and 0.01

mIU/mL

+

0.01

respectively

(see Table 4).

Table

4.

Concentration ofthe FSH and LH hormones before,

during, and after TE and DMPA injection

Weeks Gonadotropin hormones (n = 6)

FSH (mIU/mL) +

SD

LH (mIU/mL) + SD

Med J lndones

volume

is

in

line with

diameter

of

seminifèrous

tubules whièh

becomes

smaller and also

with

the

scores

of

spermatogenesis

which

becomes

lower.

Thus,

the decrease

in

testes

volume

indicates

a

histological

structure

disorder.

tubule

is

followed

by hyalinization

of

the basal membrane.

Progressive

hyalinization

could

be

an

inhibitor

between

peri-tubular capillaries

and

seminiferous tubule epithelium.

This condition would

accelerate

atrophy

of

germinal

cells.t2

Hyalinization was unclear, however,

that

is

common as

this

was due to hormonal

mechanism

disorder

at

the hypothalamus-hypophysis-testes

axis through

two

ways i.e.

h

m

and

hypergonadotrophi

o-trophic

hypogonadism

affects the hypothalamus

and

hypophysis, while

hypergonadotrophic hypogonadism

was an expression

of

tubular

disgenesis. This

study

shows

that

damage

on histological structure

were

hyalinization

of seminiferous tubule

basale membrafle and spermatogenesis

disorder

is

demonstrated

by low

spermatogenesis score due

to the effect

of

a

series

of

combination

of

TE and

DMPA

injection

through

hypothalamus-hypophysis-testes

hormonal

axis'

In

relation to

hormone

levels

in

the serum, the level of

free testosterone

and estradiol

increased

during TE

and DMPA injection. This could force the

feedback

system

at

hypothalamus

and

hypophysis,

and

influence

secretion

of FSH

and

LH

hormone.

Results

of this study

are also

in line with Sutyarso's study

on

cynomolgus

macaques,

which

stated

that injection of

combination

of TE

and

DMPA

will

increase

free

testosterone hormone in serum.

This

condition wrll

also

result

on

secretion

of

LH

and FSH by

hypophysis.r3

Therefore,

the

injection

of

TE

and

DMPA

has a

direct effect on

LH

and

FSH levels

and an indirect

effect

on testes

function

disorder.

The

TE

influence

on

reproductive activities in

the

bodv

will

stay for 1.5 months while

DMPA,

for

3

months.'t fne

fg

and

DMPA injection did not

have

influence

after that

period. Therefore after week 30,

it

is considered

as

the recovery

period. Data show

that

testes

volume, diameter

of

semiferous tubule,

and

score

of

spermatogenesis increase

after week

30.

This

indicates that influence on

both

spermatogenesis and

hormonal reproduction

levels

is

reversible'

This

happened

to

bqqh

cynomolgus macaquesr3

and

volunteers (men).''o

-t2

0

6

T2

t8 24 30 36

42

48

3.06 + 1.06 2.44 + 1.33 0.26 + 0. l3

0.28 + 0.18

2.26 + 2.72 2.41 -r 2.18

4.Ol + 4.67

4.91 + 6.48

2.99 + 4.33

1.1

I

+ 2.16

0. 12 + 0.06 0.10 + 0.00

0.00 + 0.00

0.07 + 0.10

0.09 + 0.12

0.05 + 0.09 0.04 + 0.07

0.00 + 0.00 0.01 + 0.03 0.01 + 0.01

DISCUSSION

Results

of

the study on body weight of pig

tail

macaques

indicate that the feedback system

disorder

of

hypo-thalamus-hypophysis-testes could not be demonstrated

by

only

measuring

their body weight.

Suhana et al.(1999)

reported

that

the two

groups

of

cynomolgus macaques

(Macaca

fascicularis) that were injected

with

com-bination of TE

and

DMPA

and were fed

with different

concentrations

of

fat

and

protein resulted in different

body weights. This difference, however,

ha.s^no

effect

on the feedback mechanism

of both

grciups.r0

Results

indicate that

there

is

a relationship

between

injection of TE

ànd

DMPA

and testes

volume.

Study on measurement

of seminiferous tubules

diameter and

scores

of

spermatogenesis

shows

that

decrease of

testes volume during injections

of

TE

and DMPA

[image:6.612.46.274.303.468.2]

(7)

CONCLUSIONS

Injection

of

TE

32

mg

and

DMPA

40 mg can decrease

testes

volume

and influence testes

histological

structure,

i.e.

decreased diameter

of

semiuiferous tubules, basale

membrane

hyalinization

and

causes spermatogenesis

inhibition.

This

injection also directly

increase

the

level

of

total

testosterone,

free

testosterone,

and

estradiol

in

serum,

and also

decrease serum

FSH

and

LH.

The

influence

of

injection

of

TE

and DMPA

combination

on

spermatogenesis

and hormonal

re-production levels

are reversible.

Acknowledgement

We thank the

government

of

Indonesia (Directorate

General of

Higher

Education, Department

of

National

Education)

for

supporting this research under the Project

of

Research

Team

'URGE'

No

023/HTPPruRGE/1995.

We

are

grateful to Dr. Dondin

Sayuti, director

of

the

Primate

Research

Center and

his staff

for

providing

the facilities required

for

undertaking

this

research.

We are

especially grateful

to

veterinarians

Dr.

I.

Nengah Budiarsa and

Dr.

Ivov Rinaldi

Hasibuan

for

their

guidance

and

help

in

handling

of

the non-human primates as research animals.

REFERENCES

1.

Bremmer WJ,

De

Kretser

DM.

The prospects

for

new,

reversible male contraceptives. New Eng

J

Med. 1976;

1 l:1 111-7 .

2.

Wu FCW. Male contraception. Current status and future

prospects. Clin Endocrin 1988; 29:443 -65.

Matsumoto

AM.

Is high dosage testosterone an effective

male agent? Fertil Steril. 1988; 50:324-8.

World

Health

Organization

(WHO).

Contraceptive

efficacy

of

testosterone induced azoospermia

in

normal

men. Lancet 1990; 336: 955-9.

Knuth UA, Yeung CH, Nieschlag E. Combination

of

19-NTHPP and DMPA for male contraception. Fertil Steril.

1989;51:1011-8.

Pangkahila

W.

Reversible azoospermia

induced

by

androgen progestin combination regimen

in

Indonesia

men. Int J Androl 1991; 14:248-56.

Moeloek

N.

Comparison

of

two androgens plus DMPA

for

suppression to azoospermia

in

Indonesia men. Fertil

Steril. 1993; 60: 1062-8.

Bercovitch

FB,

Rodriguez

JF.

Testis size, epididymis

weight, and sperm competition in rhesus macaques. Am J

Primat 1993; 30:163-8.

Johnsen SG. Testicular biopsy score count-a method

foi

registration

of

spermatogenesis

in

human testis: normal

values and results

in

335 hypogonadal males. Hormonqs

1970;1:1-24.

Suhana

N,

Sutyarso, Moeloek

N,

Soeradi O, Sukmaniah

SS, Supriatna J. The effects of feeding afr Asia or Western

diet on sperm number, sperm quality and seruln hormone

levels

in

_{cynomolgus monkeys (Macaca fascicularis)} injected

with

testosterone enanthate

(TE)

plus

depot

medroxyprogesterone acetate (DMPA). Int J Androl 1999;

22:102-12.

Marson J, Meuris, Cooper RW, Jouannet P. Puberty in the

male

Chimpanzee: progressive maturation

of

semen

characteri sti cs. Biol Reprod 199 1 ; 44:446-5 5.

Nistal M, Paniagua R. Testicular and epididymal pathology.

New York: Thieme-stratton Inc. 1984.

Sutyarso. Pengaruh pemberian pakan berkadar protein,

lemak,

dan

karbohidrat berbeda terhadap timbulnya

azoospermia pada monyet jantan (Macaca fascicularis)

yang disuntik kombinasi testosterone enântat

(TE)

dan

depot medroksiprogesteron asetat (DMPA) [Disertation].

Jakarta: Univ of lndonesia:' 1997 .

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4

5

6

8

9

t0

11.

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